This Test Guideline describes an in vitro screen for chemical effects on steroidogenesis,
specifically the production of 17ß-estradiol (E2) and testosterone (T). The human
H295R adreno-carcinoma cell line, used for the assay, expresses genes that encode
for all the key enzymes for steroidogenesis. After an acclimation period of 24 h in
multi-well plates, cells are exposed for 48 h to seven concentrations of the test
chemical in at least triplicate. Solvent and a known inhibitor and inducer of hormone
production are run at a fixed concentration as negative and positive controls. At
the end of the exposure period, cell viability in each well is analyzed. Concentrations
of hormones in the medium can be measured using a variety of methods including commercially
available hormone measurement kits and/or instrumental techniques such as liquid chromatography-mass
spectrometry. Data are expressed as fold change relative to the solvent control and
the Lowest-Observed-Effect-Concentration. If the assay is negative, the highest concentration
tested is reported as the No-Observed-Effect-Concentration.
Published on July 04, 2023Also available in: French
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